新藤黄酸干预肺腺癌血管生成的研究

［］目的 通过对人脐静脉内皮细胞HUVEC和人肺腺癌A549的培养,检测含新藤黄酸(gambogenic acid,GNA)条件培养基对血管内皮细胞存活率、成管和生长的影响。方法 采用甲基噻唑基四唑(methyl thiazolyltetrazolium,MTT)法和平板克隆实验法研究新藤黄酸对HUVEC存活率和克隆形成率的影响;应用薄层胶原建立血管内皮细胞的二维培养模型,观察新藤黄酸对于血管内皮细胞成管现象的影响;采用细胞划痕愈合和小室迁移实验考察新藤黄酸对 HUVEC的迁移能力影响;Western-blotting检测血管内皮生长因子(VEGF)和缺氧诱导因子(HIF-1α)蛋白的表达。结果 MTT检测结果显示,HUVEC细胞存活率和克隆形成率随GNA剂量增加而降低。新藤黄酸可抑制HUVEC细胞的迁移。还可抑制HUVEC管腔样结构形成。此外,新藤黄酸可下调HUVEC中VEGF和HIF-1α蛋白的表达。结论 新藤黄酸可在体外抑制血管生成,其作用机制可能与抑制肿瘤细胞分泌的缺氧诱导因子(HIF-1α)和血管内皮生长因子(VEGF)有关。     

蜈蚣藻多糖的体外抗血管生成作用

目的探讨蜈蚣藻多糖(GFP)的体外抗血管生成作用。方法应用MTT比色法测定GFP对人脐静脉内皮细胞(HUVEC)存活率的影响;用Millicell小室测定GFP对HUVEC迁移能力的影响;用Matrigel测定GFP对HUVEC管腔形成的影响。结果GFP在5、10、20、50、100mg.L-1浓度下处理HUVEC48h,细胞存活率没有显著性变化;在50、100mg.L-1时能显著性地抑制HUVEC的迁移并呈剂量依赖关系;在100mg.L-1时能显著性地抑制HUVEC的管腔形成。结论GFP能够抑制HUVEC的迁移和管腔的形成。     

西妥昔单抗抑制血管生成的实验研究

目的 探讨西妥昔单抗(C225)对体内、外血管生成的抑制作用.方法 用人脐静脉内皮细胞(HUVEC)原代培养模型,分别采用四甲基偶氮唑盐(MTT)法、Transwell小室趋化实验、体外小管形成实验及流式细胞术,观察C225在体外对HUVEC增殖、迁移、小管形成和凋亡的影响;利用鸡胚绒毛尿囊膜(CAM)模型,观察C225对CAM新生血管的抑制作用.结果 0.0625～2.0000μg/ml的C225作用HUVEC 48 h,细胞增殖抑制牢为12.34%～25.26%;体外迁移及小管形成实验显示,0.0625～1.0000 μg/ml C225的HUVEC迁移抑制率为15.51%～48.68%,HUVEC小管形成抑制率为22.91%～44.71%;0.25 μg/ml和1.00 μg/ml的C225诱导HUVEC细胞凋亡率分别为33.20%和35.05%.体内CAM试验表明,0.0625～1.0000 μg/ml的C225对新生血管抑制率为20.39%～36.18%.结论 C225在体内、外具有血管生成作用.     

毛萼乙素抑制人正常及肝癌血管生成的作用研究

目的:研究毛萼乙素对人脐静脉内皮细胞(HUVEC)增殖的影响以及对人肝癌血管生成的影响。方法:采用改良MTT法测试6个浓度的毛萼乙素在体外对原代培养的HUVEC增殖的影响;用人肝癌鸡胚尿囊膜血管复制模型,测定毛萼乙素在器官水平对血管生成的抑制作用。结果:毛萼乙素对HUVEC增殖具有显著的抑制作用,半数抑制浓度(IC50)=7.27μg·mL-1;抑制效应呈剂量和时间依赖性。在剂量为25、50、100μg·mL-1时对人肝癌鸡胚尿囊膜血管面积比的抑制率分别为14.15%、48.72%、60.63%,抑制作用中等;其中剂量在50μg·mL-1时与阳性对照反应停相当(P>0.05)。结论:毛萼乙素对人脐静脉内皮细胞的增殖有显著的抑制作用,并能显著抑制人肝癌细胞诱导的鸡胚尿囊膜血管生成。     

siRNAs对原代人脐静脉内皮细胞血管生成的影响

目的 研究siRNA单独或多重沉默原代人脐静脉内皮细胞(HUVECs)中血管内皮细胞生长因子A(VEGFA)及血管内皮细胞生长因子受体1/2/3(VEGFR1/2/3)表达对其血管生成的影响。方法 HUVECs分为空白对照组,阴性对照组与实验组。空白对照组不给予转染,阴性对照组转染100 nmol·L^-1siNC,实验组分为4组并分别转染100 nmol·L^-1siVEGFA及siVEGFR1/2/3。以反转录-聚合酶链反应(RT-PCR)检测siRNAs的沉默效率。HUVECs分为空白对照组,阴性对照组,沉默单靶点组和多靶点组,空白对照组不给予转染,阴性对照组转染100 nmol·L^-1siNC,沉默单靶点组分为4组并分别转染100 nmol·L^-1siVEGFA及siVEGFR1/2/3,沉默多靶点组分为4组并分别转染100 nmol·L^-1siVEGFA+siVEGFR1、siVEGFA+siVEGFR2、siVEGFA+siVEGFR3及siVEGFR1+siVEGFR2+siVEGFR3,以小管形成实验检测各组HUVECs小管形成情况。结果 siVEGFA组、siVEGFR1组、siVEGFR2组与siVEGFR3组沉默效率分别为(85.67±10.50)%,(71.67±7.02)%,(89.00±11.36)%与(90.00±3.60)%,与空白及阴性对照组相比,差异均有统计学意义(均P<0.01)。siVEGFA组血管总长度、分支管长度和血管交叉点分别为(32.67±3.79)%,(22.67±2.08)%及(25.33±6.43)%,在沉默单靶点组中抑制血管生成能力最强(均P<0.05);siVEGFA+siVEGFR3组血管总长度、分支管长度和血管交叉点分别为(28.33±5.51)%,(10.00±2.65)%,(8.33±2.31)%,siVEGFR1+siVEGFR2+siVEGFR3组分别为(27.00±4.00)%,(9.33±2.89)%,(10.00±3.60)%,在沉默多靶点组中抑制血管生成能力最强(均P<0.05)。结论 siRNAs沉默VEGF通路中单靶点的表达,可有效抑制HUVECs诱导的血管生成;同时沉默多靶点的表达,其抑制血管生成的功效更强。     

新绿原酸抑制血管生成的研究

目的 考察新绿原酸(5-caffeoylquinic acid, 5-CQA)对血管生成的影响。方法 应用人脐内皮细胞(HUVEC)、大鼠动脉环模型和转基因血管荧光斑马鱼模型考察5-CQA对血管生成的抑制作用。同时建立肿瘤细胞斑马鱼移植瘤模型,考察5-CQA对肠下静脉(subintestinal vein, SIV)生成的作用。结果 5-CQA在320μmol·L^-1及以下浓度对HUVEC的增殖无明显影响,5-CQA能够抑制HUVEC体外迁移及细胞中VEGF、bFGF mRNA的表达;与对照组比较,56和112μmol·L^-1的5-CQA具有显著抑制大鼠动脉环微血管生成的作用(P<0.05);在斑马鱼血管荧光模型中56和112μmol·L^-1的5-CQA具有显著抑制24 h斑马鱼节间血管(intersegmental vessel, ISV)生成的作用;同时5-CQA对斑马鱼移植瘤的肠下静脉生成具有明显抑制作用(P<0.05)。结论 5-CQA具有抑制血管生成的作用,其作用机制与抑制VEGFR2受体的表达有关...     

血管生成抑制剂研究进展

目的介绍近年来血管生成抑制剂的研究进展。方法查阅相关文献进行总结、归纳。结果血管生成抑制剂的研究取得很大进展。结论对寻找高效、低毒副作用的血管生成抑制剂具有重要意义。     

血管生成抑制剂研究进展

目的 介绍近年来血管生成抑制剂的研究进展。     

参麦注射液增强羟基喜树碱对肿瘤血管生成的抑制作用研究

目的研究参麦注射液联合羟基喜树碱对肿瘤血管生成的影响.方法:将人结肠癌细胞株(CW-2)细胞分为4组,即空白、参麦、羟基喜树碱、联合用药组.分组后的CW-2细胞培养12 h,制得条件培养液,条件培养人脐静脉内皮细胞(HUVEC),MTT法检测细胞生存率,并用细胞培养小室检测其细胞迁移活性.结果:与空白组比较,其他3个用...     

鬼箭羽提取物抑制血管生成的实验研究

目的 研究鬼箭羽氯仿提取物对血管生成的影响.方法 采用人脐静脉内皮细胞模型（HUVEC）、大鼠动脉环模型（rat aortic rings）和鸡胚绒毛尿囊膜（CAM）模型研究鬼箭羽提取物抑制血管生成作用.结果 8μg·mL-1鬼箭羽提取物显著抑制人脐静脉内皮细胞的增殖,抑制率达到36.2%;2μg·mL-1和4μg·mL-1鬼箭羽提取物能够显著抑制大鼠动脉环新血管结构形成,抑制率分别达到56.41%和65.25%;鬼箭羽提取物20 μg·个-1与40μg·个-1.对鸡胚绒毛尿囊膜（CAM）血管抑制率分别达到了22.6%与31.2%.结论 鬼箭羽提取物具有显著的抗血管生成活性.     

Arsenite suppresses angiogenesis of vascular endothelial cells mediated by Platelet Derived Growth Factor Receptor-beta

The present study aimed to investigate the effects of sodium arsenite (NaAsO₂) on the angiogenesis of human umbilical vein endothelial cells (HUVECs) and the mechanism involved. Firstly, a Matrigel-based in vitro angiogenesis assay demonstrated that arsenite suppressed the angiogenesis of HUVECs in a dose-dependent manner. Then by using a global inhibitor for multiple growth factor receptors (E7080) and a specific inhibitor of PDGFR-beta (CP-673451), we found that E7080 completely prevented and CP-673451 significantly decreased the angiogenesis of HUVECs. This suggested that angiogenesis of HUVECs depends on the signal pathway mediated by tyrosine kinase receptors and that among them, PDGFR-beta has an important regulatory function. Finally by using porcine aortic endothelial cells which stably express human PDGFR-beta, we found that arsenite suppressed the angiogenesis mediated by PDGFR-beta. Based on these results, we conclude that arsenite suppressed the angiogenesis of the vascular endothelial cells, that this effect is mediated by PDGFR-beta, and postulate that it might contribute to the injuries of blood vessel in arsenism.     

阿托伐他汀对血管新生的促进作用

目的探讨阿托伐他汀对血管新生的促进作用及其作用机制。方法建立野生型C3H／He小鼠下肢缺血模型,观察用药后肢体血流、毛细血管数、血管内皮生长因子（VEGF）蛋白表达。并行离体实验（血管新生共同培养）,计数血管样结构物管腔形成数。结果阿托伐他汀使实验小鼠术后缺血肢血流明显改善,缺血肢与非缺血肢血流面积比明显增加;缺血肢毛细血管密度明显增加,VEGF蛋白表达增强。血管新生共同培养,血管样结构物管腔数明显高于对照组和加入血管新生抑制剂组,但与阳性对照组比较无明显差异。结论阿托伐他汀有促进血管新生的作用。     

Zerumbone,Sesquiterpene Photochemical from Ginger,Inhibits Angiogenesis

Here, we investigated the role of zerumbone, a natural cyclic sesquiterpene of Zingiber zerumbet Smith, on angiogenesis using human umbilical vein endothelial cells (HUVECs). Zerumbone inhibited HUVECs proliferation, migration and tubule formation, as well as angiogenic activity by rat aorta explants. In particular, zerumbone inhibited phosphorylation of vascular endothelial growth factor receptor-2 and fibroblast growth factor receptor-1, which are key regulators of endothelial cell function and angiogenesis. In vivo matrigel plug assay in mice demonstrated significant decrease in vascularization and hemoglobin content in the plugs from zerumbone-treated mice, compared with control mice. Overall, these results suggest that zerumbone inhibits various attributes of angiogenesis, which might contribute to its reported antitumor effects.     

血管内皮生长因子在非小细胞肺癌中的表达

目的 探讨血管内皮生长因子(VEGF)在非小细胞肺癌(NSCLC)中的表达及临床意义.方法 应用免疫组化法检测100例根治术后且术前未接受过放化疗等抗癌治疗的NSCLC患者的癌组织及正常癌旁组织中VEGF的表达.结果 VEGF在癌组织中的表达明显高于在癌旁正常组织(P＜0.01).癌组织中VEGF的表达与性别、年龄、血型、病理类型、TNM分期无明显相关性(P＞0.05).结论 VEGF在癌组织中的表达增高,提示VEGF在NSCLC的发生发展中具有促进作用.     

Antiangiogenic effect of licochalcone A

To date, no antiangiogenic activity has been demonstrated for licochalcone A (LicA), a major phenolic constituent of Glycyrrhiza inflata, although it shows significant antitumor activity in human malignant cell lines. Our previous work demonstrated that LicA down-regulates inflammatory responses to lipopolysaccharide in murine macrophages. The purpose of the present study was to evaluate whether LicA inhibits angiogenesis, which is crucial for cancer development and progression. LicA significantly inhibited proliferation (20 μM), migration (5-20 μM), and tube formation (10-20 μM) of human umbilical vascular endothelial cells (HUVECs) as well as microvessel growth from rat aortic rings (10-20 μM). Furthermore, LicA significantly inhibited the growth of CT-26 colon cancer implants in BALB/c mice, with fewer CD31- and Ki-67-positive cells but more apoptotic cells. The underlying antiangiogenic mechanism of LicA correlated with down-regulation of vascular endothelial growth factor receptor (VEGFR)-2 activation. Our findings provide the first evidence that LicA inhibits angiogenesis in vitro and in vivo, perhaps by blocking VEGF/VEGFR-2 signaling. Inhibition of tumor growth may be attributed, at least in part, to decreased angiogenesis in LicA-treated mice. These findings emphasize the potential use of LicA against tumor development and progression in which angiogenesis is stimulated.     

Pseudolarix acid B inhibits angiogenesis by antagonizing the vascular endothelial growth factor-mediated anti-apoptotic effect

Angiogenesis is controlled by a number of growth factors, including vascular endothelial growth factor (VEGF). In this study, pseudolarix acid B, isolated from the traditional Chinese medicinal plant Pseudolarix kaempferi and originally identified as an early pregnancy-terminating agent, was evaluated for its potential as an angiogenesis inhibitor, using in vitro and in vivo models. After exposure to pseudolarix acid B 0.625-5 microM for 72 h, the proliferation of human umbilical vein endothelial cells was significantly inhibited. Pseudolarix acid B 0.313-2.5 microM for 24 h potently blocked the VEGF-induced tube formation of human umbilical vein endothelial cells in a dose-dependent manner. Matrigel plug assays disclosed that pseudolarix acid B reduced angiogenesis induced by VEGF in vivo. In addition, pseudolarix acid B antagonized VEGF-mediated anti-apoptotic effects on serum-deprived human umbilical vein endothelial cells and increased apoptosis of endothelial cells induced by VEGF in Matrigel plug assays. Moreover, pseudolarix acid B significantly inhibited VEGF-induced tyrosine phosphorylation of kinase insert domain-containing receptor/fetal liver kinase-1 (KDR/flk-1), in correlation with a marked decrease in the phosphorylation of Akt and extracellular signal-regulated kinases (ERK). These findings collectively suggest that pseudolarix acid B possesses anti-angiogenic activity. One of the main anti-angiogenesis mechanisms of pseudolarix acid B may involve antagonism of the VEGF-mediated anti-apoptosis effect via inhibition of KDR/flk-1, ERK1/2, and Akt phosphorylation in endothelial cells.     

The effects of cadmium on VEGF-mediated angiogenesis in HUVECs

Cadmium (Cd) is a highly toxic element that causes morphologic alterations and dysfunction in blood vessels. The altered vascular function caused by cadmium has been implicated in a range of chronic diseases, including hypertension. The effects of cadmium are a multisystem phenomenon involving inflammation, hypertrophy, apoptosis, angiogenesis and important processes involved in vascular remodeling systems. Vascular endothelial growth factor (VEGF) plays a major role in cell growth and angiogenesis under pathologic conditions. VEGF secretion is related to anti-apoptosis protein expression and attenuates apoptosis in endothelial cells. This study examined the VEGF-dependent mechanisms of angiogenesis and apoptosis in cadmium-treated endothelial cells (HUVECs). The effects and mechanisms of cadmium in endothelial cells (HUVECs) were examined by exposing the cells to different doses of cadmium chloride (2.5-40 μ m). After the cadmium treatment, the angiogenesis and apoptosis mechanisms related to VEGF in cadmium-treated HUVECs were examined. As a result, the low concentration of cadmium increased the tube formation in HUVECs. In addition, cadmium at concentrations of 5 and 10 μ m increased VEGF secretion and VEGFR2 activity, which suggest that cadmium affects the growth of blood vessels. All three MAPK pathways, namely ERK, JNK and p38, were activated by cadmium in HUVECs. However, high concentrations of cadmium caused cell damage, disrupted tube formation and inhibited VEGF expression and the activities of VEGFR2 and MAPK in HUVECs. Cadmium has dual functions through VEGF-dependent mechanisms in a dose-dependent manner. In this study, the dual effects of cadmium might alter angiogenesis and induce apoptosis through VEGF pathways in HUVECs.     

乌司他丁对脂多糖诱发的单层人脐静脉内皮细胞高通透性的影响及其机制

目的 探讨乌司他丁对脂多糖(LPS)诱发的单层人脐静脉内皮细胞(HUVECs)高通透性的影响及其可能机制.方法 体外培养的单层 HUVECs被分为对照组、LPS组(LPS 1μg/ml刺激4 h)、乌司他丁复合LPS组(乌司他丁3000 U/ml孵育1 h+LPS 1μg/ml刺激4 h)和血管内皮钙黏蛋白单克隆抗体(VE-cadherin mAb)组(VE-cadherin mAb 50μg/ml孵育6 h+乌司他丁3000 U/ml孵育1 h+LPS 1μg/ml刺激4 h).采用双腔系统Transwell法检测单层 HUVECs的通透性 , Western blot法检测HUVECs中VE-cadherin的表达水平.结果 与对照组相比,LPS组单层HUVECs渗透性增高 ,VE-cadherin表达降低(P＜0 .01).与LPS组相比 ,乌司他丁复合LPS组单层HUVECs渗透性降低 ,VE-cadherin表达增高(P＜0 .01).与乌司他丁复合LPS组相比 ,VE-cadherin mAb组单层HUVECs渗透性增高 ,VE-cadherin表达降低(P＜0 .01).结论 乌司他丁预处理可改善LPS引起的单层HUVECs高通透性状况 ;其可能机制与VE-cadherin表达增多有关.     

VEGF-C对宫颈癌凋亡影响的研究进展

血管内皮生长因子-C（VEGF-C）是一类很重要的淋巴管生成因子,其在宫颈癌淋巴管转移方面的研究较多,也证明了VEGF-C在宫颈癌发生发展、侵袭转移中起重要作用。然而,VEGF-C在宫颈癌中的作用机制并未完全明确,其对宫颈癌凋亡的影响值得继续探索。本文就VEGF-C在宫颈癌凋亡中的作用及其机制进行综述。